基因免疫制备牛分枝杆菌MPB64抗原单克隆抗体  

Preparation of monoclonal antibodies against the MPB64 antigen of Mycobacterium bovis by gene immunization

作　　者：刘建东[1] 刘思国[1] 王春来[1] 宫强[1] 王勇[2] 迟磊[1] 赵昆[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所国家兽医生物技术重点实验室/动物细菌病研究室,黑龙江哈尔滨150001 [2]东北农业大学,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2007年第11期870-873,共4页Chinese Journal of Preventive Veterinary Medicine

摘　　要：为了建立针对牛分枝杆菌分泌抗原MPB64的检测手段,制备该抗原的单克隆抗体,构建真核表达载体pcDNA-mpb64,以原核表达并纯化的融合蛋白H-MPB64包被ELISA板,建立单克隆抗体检测方法。利用基因免疫方法免疫BALB/c小鼠,通过杂交瘤技术筛选出针对牛分枝杆菌分泌蛋白MPB64的单克隆抗体一株,并制备了腹水,经鉴定腹水效价达到1∶10240,采用硫酸铵沉淀法纯化腹水,纯化抗体效价为1∶8000,为今后研制分枝杆菌鉴定技术奠定基础。To establish a new detection method for MPB64 antigen of M.bovis, a mammalian expression vector pcDNAmpb64 was constructed and used to immunize the BALB/c mice. A prokaryotic system expressing H-MPB64 protein as detective antigen was used for testing monoclonal antibodies against the MPB64 antigen of M.bovis. One hybridoma cell line secreting monoclonal antibodies was established. The McAb reacted with H-MPB64 and PPD in ELISA experiment, but not in western blot.

关 键 词：牛分枝杆菌MPB64蛋白 基因免疫 单克隆抗体 

分 类 号：S852.43[农业科学—基础兽医学]